Growth of T-lymphoma cells in serum-free medium: lack of involvement of the cyclic AMP pathway in long-term cultures.

We have developed a serum-free, chemically defined growth medium containing casein, insulin, transferrin, testosterone, and linoleic acid in Dulbecco's modified Eagle's medium/Ham's F12 medium, 1:1 (vol/vol), for growing murine T lymphomas. This medium supports the growth in suspension of all murine T lymphomas tested, including S49, WEHI 7, EL4, BW5147, and R1.1. Growth of these cell lines was maintained indefinitely with doubling times approaching those of cells grown in 10% (vol/vol) horse serum. This medium also supports the growth of several of the S49 variants of the beta-adrenergic receptor/adenylate cyclase/cyclic AMP/protein kinase pathway, suggeting little or no involvement of this pathway in the routine growth of S49 cells or in the mechanism of action of the factors in this defined medium. This serum-free medium should prove useful for studies of a variety of metabolic pathways and of differentiated functions of T-lymphoma cells.